LOCUS       sequence-301680-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6804)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6804)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4252945
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    63..184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(100..119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     154..173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(253..270)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(271..291)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(591..793)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(620..640)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        795..1098
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1179..1227
                     /note="synthetic polyadenylation signal"
     terminator      1245..1291
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1296..1367
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1519..1655)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1668..2459,2460..2528))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2291..2310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2794..2842
                     /note="synthetic polyadenylation signal"
     CDS             complement(2869..3897)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3901..5110)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4045..4064)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5610..6198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6099..6118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6424..6495
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6540..6676)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6706..6752)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
       61 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      121 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      181 cttatcatgt ctgtgagctg aaggtacgct gtatctcagt cagtcaagct agcagaggcc
      241 tgcggatcct taatggtgat ggtgatggtg cgactgaaag taaaggtttt cgacaagaag
      301 atcaggcaca cgggagctgt ttagattttt cacgcgggaa taaacgtaga acgagggctt
      361 cacgagacta acgttgacaa tgttacaaca ataagcgcac agccggaggg ccgtcagtat
      421 cgctagggtc actaaaagga agaccacaaa tgccaggaac agcaacactg aattaacgat
      481 gagagtgcct gtctcttcag atacaaagct gtacatggtg gcggccgctg gcgggtgtgt
      541 cgagtggatg gtaggatcga caaagatctg atctgacggt tcactaaacc agctctgctt
      601 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta
      661 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
      721 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc
      781 caaaaccgca tcagcatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
      841 aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
      901 tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
      961 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     1021 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     1081 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     1141 gattactaga tctggctaca ctcgatcagc agttagataa taaaatcgct atccatcgaa
     1201 gatggatgtg tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg
     1261 ggcctctaaa cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat
     1321 gaccactaga gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga
     1381 tcgctgccaa aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg
     1441 agcgccacgg ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag
     1501 tttaaacgat tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
     1561 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
     1621 agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta
     1681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1801 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt
     1921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt
     1981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2101 ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa
     2221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag
     2341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2401 cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2521 atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg
     2581 actcgattat acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc
     2641 tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga
     2701 ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc
     2761 gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc
     2821 ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagctt attcctttgc
     2881 cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc
     2941 ggtccagacg gcggcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga
     3001 tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac
     3061 caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga gccggggcga
     3121 tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa
     3181 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
     3241 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
     3301 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
     3361 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
     3421 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
     3481 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gcggcagcga tggcatccat
     3541 tgcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
     3601 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc
     3661 aagcacttcc ggaatcggga gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt
     3721 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
     3781 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
     3841 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tacccatgtt
     3901 gtctaacaaa aaagccaaaa acggccagaa tttagcggac aatttacaag tctaacactg
     3961 aaaattacat attgacccaa atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa
     4021 cacactcgcc aaccccgagc gcatagttca aaaccggagc ttcagctact taagaagata
     4081 ggtacataaa accgaccaaa gaaactgacg cctcacttat ccctcccctc accagaggtc
     4141 cggcgcctgt cgattcagga gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag
     4201 aaaagcctac cgcacaccta ccggctggtg gccccaccct gcattataag ccaacagaac
     4261 gggtgacgtc acgacacgac gagggcgcgc gctcccaaag gtacgggtgc actgcccaac
     4321 ggcaccgcca taactgccgc ccccgcaaca gacgacaaac cgagttctcc agtcagtgac
     4381 aaacttcacg tcagggtccc cagatggtgc ccagcccatc tcacccgaat aagagctttc
     4441 ccgcattagc gaagggctca agaccttggg ttcttgccgc ccaccatgcc ccccaccttg
     4501 tttcaacgac ctcacagccc gcctcacaag cgtcttccat tcaagactcg ggaacagccg
     4561 ccattttgct gcgctccccc caacccccag ttcagggcaa ccttgctcgc ggacccagac
     4621 tacagccctt ggcggtctct ccacacgctt ccgtcccacc gagcggcccg gcggccacga
     4681 aagccccggc cagcccagca gcccgctact caccaagtga cgatcacagc gatccacaaa
     4741 caagaaccgc gacccaaatc ccggctgcga cggaactagc tgtgccacac ccggcgcgtc
     4801 cttatataat catcggcgtt caccgcccca cggagatccc tccgcagaat cgccgagaag
     4861 ggactacttt tcctcgcctg ttccgctctc tggaaagaaa accagtgccc tagagtcacc
     4921 caagtcccgt cctaaaatgt ccttctgctg atactggggt tctaaggccg agtcttatga
     4981 gcagcgggcc gctgtcctga gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg
     5041 tctgacgtgg cagcgctcgc cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg
     5101 cgcggaggcc aggcgcgccc cgatttcgat attcctcgga tctcggatag atcgggattg
     5161 aggatgtagg ttgaacttct ctcgggtgat cccgatctcg gatattcggg attggagtaa
     5221 tacctggcga taggcgctgc caagcaatag ccaaggcgca aggccggcca agagacttgc
     5281 gatagttcgt tcgagaggta gcctgcaaga tggatcggac gaaaggtttg aagattttcc
     5341 gacgagggcc ttggagagtt cggaaattta aatgcaacta tggatgaacg aaatagacag
     5401 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5461 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5521 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6001 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6241 tggccttttg ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa
     6301 ggaccagatc gaacttgact gacgcaattc tgagcgtagt cgaaacccag acaagtcgat
     6361 ctcgccaagt agctgacttg actggttaat taagtttcga tagcccaagg taaccaatga
     6421 gattagggag ggaagaaagt ttgatgaact acttctgacc cctagtggtg tccagaaaag
     6481 accattaaag gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag ctgacgacct
     6541 taaggataaa tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta
     6601 tccgttgctg aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt
     6661 ttttctgcgc caatgccggt agatagcaat ccggcccgag gggcacaaaa aacccctcaa
     6721 gacccgttta gaggccccaa ggggttatgc tacgataccg atagagatgg gccaatacca
     6781 catttgtaga ggttttactt gctt
//
